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DNA Isolation

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Rapid, Safe, and Simple Manual Bedside Nucleic Acid Extraction for the Detection of Virus in Whole Blood Samples

Rapid, Safe, and Simple Manual Bedside Nucleic Acid Extraction for the Detection of Virus in Whole Blood Samples

Published on: June 30, 2018

Extractionless nucleic acid detection: a high capacity solution to COVID-19 testing.

Shairaz Baksh¹, Natalia Volodko², Merle Soucie²

¹DynaLIFE Medical Labs, Edmonton, Alberta, Canada; BioImmuno Designs, Edmonton, Alberta, Canada.

Diagnostic Microbiology and Infectious Disease

|July 18, 2021

Summary

This summary is machine-generated.

A new heat-based, extractionless protocol for SARS-CoV-2 detection using real-time reverse transcriptase-PCR (rRT-PCR) offers accurate, cost-effective COVID-19 testing. This method significantly increases testing capacity and reduces turnaround time for results.

Keywords:

COBAS 4800 COVID-19 E-gene Extractionless Heat extraction N1-gene N2-gene Orf1ab gene Quantabio RdRp gene Roche SARS-CoV-2

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Filtration Isolation of Nucleic Acids: A Simple and Rapid DNA Extraction Method

Filtration Isolation of Nucleic Acids: A Simple and Rapid DNA Extraction Method

Published on: August 6, 2016

Area of Science:

Molecular Biology
Virology
Clinical Diagnostics

Background:

Accurate and rapid detection of SARS-CoV-2 is crucial for controlling the COVID-19 pandemic.
Traditional nucleic acid extraction methods are time-consuming and require specialized equipment, limiting testing capacity.
Development of simplified, high-throughput diagnostic protocols is essential.

Purpose of the Study:

To develop and validate an extractionless real-time reverse transcriptase-PCR (rRT-PCR) protocol for SARS-CoV-2 detection.
To evaluate the impact of temperature, transport media, mastermixes, and gene assays on assay performance.
To assess the sensitivity, specificity, and capacity of the developed protocol for COVID-19 testing.

Main Methods:

An extractionless rRT-PCR protocol utilizing heat for SARS-CoV-2 nucleic acid detection was developed.
The effects of temperature, transport media, rRT-PCR mastermixes, and gene assays (E, N1, Orf1ab) on amplification and limit of detection were investigated.
Singleplex and multiplex (E/N1, Orf1ab/N1) assay combinations were tested.

Main Results:

The heated methodology achieved limits of detection of 12.5 genome copies/reaction for singleplex E-gene and 1 genome copy/reaction for singleplex N1-gene assays.
Multiplex assays (E/N1, Orf1ab/N1) achieved a limit of detection of 1 genome copy/reaction.
The protocol successfully detected SARS-CoV-2 in up to 98% of analyzed positive patient samples, including weak positives.

Conclusions:

The developed extractionless rRT-PCR protocol is an accurate, cost-effective, and high-capacity solution for SARS-CoV-2 detection.
This approach enables a >2-fold increase in testing capacity, eliminates the need for expensive extraction devices, and significantly reduces turnaround time.
The protocol demonstrates high sensitivity suitable for reliable COVID-19 diagnosis.